This project is to advance our knowledge of the biochemistry of complex glycosphingolipids and to improve our techniques of isolation of neutral and sulfated complex glycosphingolipids. The specific goals of the proposed research are to develop new techniques for isolation of entire populations of glycosphingolipids, to characterize the complex and tightly bound glycosphingolipids of the cell membranes and to determine which are involved in the formation of protein-lipid complexes; to isolate cell organelles and determine glycosphingolipids composition; to show the subcellular location of the enzyme(s) involved in sulfation of complex glycosphingolipids. Gastric mucosa (dog, hog) will be used to isolate complex glycosphingolipids and protein-lipid complexes. Subcellular fractionation of gastric mucosa will be performed to analyze the content and composition of glycosphingolipids in the cell organelles and to establish the location of the enzyme(s) involved in the sulfation of the complex glycosphingolipids.